Home LiteratureArticle Details
PMID: 16347576 Published · ppublish English Journal Article

A Simple and Rapid Method for Genetic Transformation of Lactic Streptococci by Electroporation.

Applied and environmental microbiology ·Vol. 54 ·No. 3 ·1988-03-00 ·Pages 655-660

Powell IB, Achen MG, Hillier AJ, Davidson BE

Abstract

An electroporation procedure for the plasmid-mediated genetic transformation of intact cells of Streptococcus cremoris and Streptococcus lactis was performed. Ten different strains were transformed. The method was simple and rapid and yielded transformant colonies in 14 to 24 h. The method was optimized for S. lactis LM0230, and transformation frequencies of between 1 x 10 and 5 x 10 transformants per mug of purified plasmid (pMU1328) were achieved routinely. The optimized procedure involved lysozyme treatment of cells. Transformation of LM0230 occurred at comparable frequencies with pLS1 (4.4 kilobase pair [kbp]), pMU1328 (7.4 kbp), and pAMbeta1 (26.5 kbp). Plasmid DNA isolated from transformants had not undergone detectable deletions or rearrangements. Transformation was possible with plasmid DNA which was religated after restriction endonuclease digestion. Phage DNA-dependent transfection of S. lactis LM0230 and S. lactis C6 was also achieved.

Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Powell Ian B
Russell Grimwade School of Biochemistry, University of Melbourne, Parkville, Victoria 3052, and Dairy Research Laboratory, Division of Food Research, Commonwealth Scientific and Industrial Research Organization, Highett, Victoria 3190, Australia.
Achen Marc G
Hillier Alan J
Davidson Barrie E
References (20)
20 references, click to expand
  1. High-efficiency transformation of Streptococcus lactis protoplasts by plasmid DNA.
    Appl Environ Microbiol. 1986 Aug;52(2):394-5 PMID: 3092737
  2. Identification and analysis of genes for tetracycline resistance and replication functions in the broad-host-range plasmid pLS1.
    J Mol Biol. 1986 Dec 20;192(4):753-65 PMID: 2438417
  3. Inorganic salts resistance associated with a lactose-fermenting plasmid in Streptococcus lactis.
    J Bacteriol. 1977 Apr;130(1):257-65 PMID: 404284
  4. Morphology of the bacteriophages of lactic streptococci.
    Appl Microbiol. 1974 Feb;27(2):411-5 PMID: 4132690
  5. A method for genetic transformation of nonprotoplasted Streptococcus lactis.
    Appl Environ Microbiol. 1987 Aug;53(8):1730-6 PMID: 3116931
  6. Physical and genetic analyses of streptococcal plasmid pAM beta 1 and cloning of its replication region.
    J Bacteriol. 1984 Feb;157(2):445-53 PMID: 6319361
  7. Enhancer-dependent expression of human kappa immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):7161-5 PMID: 6438633
  8. Improved medium for lactic streptococci and their bacteriophages.
    Appl Microbiol. 1975 Jun;29(6):807-13 PMID: 16350018
  9. Gaining access to the cytosol: the technique and some applications of electropermeabilization.
    Biochem J. 1986 Mar 15;234(3):497-506 PMID: 3521588
  10. Nucleotide sequence of the Streptococcus faecalis plasmid gene encoding the 3'5"-aminoglycoside phosphotransferase type III.
    Gene. 1983 Sep;23(3):331-41 PMID: 6313476
  11. A cloning vector able to replicate in Escherichia coli and Streptococcus sanguis.
    Gene. 1982 Oct;19(3):345-53 PMID: 6295886
  12. Electropore diameters, lifetimes, numbers, and locations in individual erythrocyte ghosts.
    FEBS Lett. 1986 Sep 15;205(2):179-84 PMID: 3743774
  13. Characterization of streptococcal bacteriophage c6A.
    J Gen Virol. 1985 Dec;66 ( Pt 12):2737-41 PMID: 2999317
  14. Plasmid complements of Streptococcus lactis NCDO 712 and other lactic streptococci after protoplast-induced curing.
    J Bacteriol. 1983 Apr;154(1):1-9 PMID: 6403500
  15. Simple and rapid method for isolating large plasmid DNA from lactic streptococci.
    Appl Environ Microbiol. 1983 Sep;46(3):549-52 PMID: 6416164
  16. Construction of plasmid vectors for the detection of streptococcal promoters.
    Gene. 1986;45(1):45-9 PMID: 3536665
  17. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  18. Plasmid transformation of Streptococcus lactis protoplasts: optimization and use in molecular cloning.
    Appl Environ Microbiol. 1984 Aug;48(2):252-9 PMID: 6091544
  19. Evidence for Plasmid Linkage of Restriction and Modification in Streptococcus cremoris KH.
    Appl Environ Microbiol. 1981 Dec;42(6):944-50 PMID: 16345908
  20. Expression of genes transferred into monocot and dicot plant cells by electroporation.
    Proc Natl Acad Sci U S A. 1985 Sep;82(17):5824-8 PMID: 3862099
Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
1098-5336
DOI
10.1128/aem.54.3.655-660.1988
Published
1988-03-00
Pages
655-660
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC202521
External Links
PubMed source DOI: 10.1128/aem.54.3.655-660.1988 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2